Identification of genetic variants associated with Huntington's disease progression: a genome-wide association study

Background Huntington's disease is caused by a CAG repeat expansion in the huntingtin gene, HTT. Age at onset has been used as a quantitative phenotype in genetic analysis looking for Huntington's disease modifiers, but is hard to define and not always available. Therefore, we aimed to generate a novel measure of disease progression and to identify genetic markers associated with this progression measure. Methods We generated a progression score on the basis of principal component analysis of prospectively acquired longitudinal changes in motor, cognitive, and imaging measures in the 218 indivduals in the TRACK-HD cohort of Huntington's disease gene mutation carriers (data collected 2008–11). We generated a parallel progression score using data from 1773 previously genotyped participants from the European Huntington's Disease Network REGISTRY study of Huntington's disease mutation carriers (data collected 2003–13). We did a genome-wide association analyses in terms of progression for 216 TRACK-HD participants and 1773 REGISTRY participants, then a meta-analysis of these results was undertaken. Findings Longitudinal motor, cognitive, and imaging scores were correlated with each other in TRACK-HD participants, justifying use of a single, cross-domain measure of disease progression in both studies. The TRACK-HD and REGISTRY progression measures were correlated with each other (r=0·674), and with age at onset (TRACK-HD, r=0·315; REGISTRY, r=0·234). The meta-analysis of progression in TRACK-HD and REGISTRY gave a genome-wide significant signal (p=1·12 × 10−10) on chromosome 5 spanning three genes: MSH3, DHFR, and MTRNR2L2. The genes in this locus were associated with progression in TRACK-HD (MSH3 p=2·94 × 10−8 DHFR p=8·37 × 10−7 MTRNR2L2 p=2·15 × 10−9) and to a lesser extent in REGISTRY (MSH3 p=9·36 × 10−4 DHFR p=8·45 × 10−4 MTRNR2L2 p=1·20 × 10−3). The lead single nucleotide polymorphism (SNP) in TRACK-HD (rs557874766) was genome-wide significant in the meta-analysis (p=1·58 × 10−8), and encodes an aminoacid change (Pro67Ala) in MSH3. In TRACK-HD, each copy of the minor allele at this SNP was associated with a 0·4 units per year (95% CI 0·16–0·66) reduction in the rate of change of the Unified Huntington's Disease Rating Scale (UHDRS) Total Motor Score, and a reduction of 0·12 units per year (95% CI 0·06–0·18) in the rate of change of UHDRS Total Functional Capacity score. These associations remained significant after adjusting for age of onset. Interpretation The multidomain progression measure in TRACK-HD was associated with a functional variant that was genome-wide significant in our meta-analysis. The association in only 216 participants implies that the progression measure is a sensitive reflection of disease burden, that the effect size at this locus is large, or both. Knockout of Msh3 reduces somatic expansion in Huntington's disease mouse models, suggesting this mechanism as an area for future therapeutic investigation

Implementación de FSSC 22000 y Reestructuración de Instalaciones en una Empresa de la Industria Gráfica del Ecuador

El proyecto por desarrollarse en esta empresa de la industria gráfica se complementa con el uso actual de materias primas menos tóxicas, nuevos controles, y medidas de reciclaje al final del proceso. Tiene como objetivo general garantizar y mantener la inocuidad de sus diferentes productos en cada una de la etapas que componen el proceso de elaboración de empaques, etiquetas y demás productos impresos sobre derivados de papel.GuayaquilMagíster en Gestón de Proyecto

Obesity modulates the immune macroenvironment associated with breast cancer development.

Growing evidence demonstrates a strong correlation between obesity and an increased risk of breast cancer, although the mechanisms involved have not been completely elucidated. Some reports have described a crosstalk between adipocytes, cancer cells, and immune cells within the tumor microenvironment, however, it is currently unknown whether obesity can promote tumor growth by inducing systemic alterations of the immune cell homeostasis in peripheral lymphoid organs and adipose tissue. Here, we used the E0771 breast cancer cell line in a mouse model of diet-induced obesity to analyze the immune subpopulations present in the tumors, visceral adipose tissue (VAT), and spleen of lean and obese mice. Our results showed a significant reduction in the frequency of infiltrating CD8+ T cells and a decreased M1/M2 macrophage ratio, indicative of the compromised anti-tumoral immune response reported in obesity. Despite not finding differences in the percentage or numbers of intratumoral Tregs, phenotypic analysis showed that they were enriched in CD39+, PD-1+ and CCR8+ cells, compared to the draining lymph nodes, confirming the highly immunosuppressive profile of infiltrating Tregs reported in established tumors. Analysis of peripheral T lymphocytes showed that tumor development in obese mice was associated to a significant increase in the percentage of peripheral Tregs, which supports the systemic immunosuppressive effect caused by the tumor. Interestingly, evaluation of immune subpopulations in the VAT showed that the characteristic increase in the M1/M2 macrophage ratio reported in obesity, was completely reversed in tumor-bearing mice, resembling the M2-polarized profile found in the microenvironment of the growing tumor. Importantly, VAT Tregs, which are commonly decreased in obese mice, were significantly increased in the presence of breast tumors and displayed significantly higher levels of Foxp3, indicating a regulatory feedback mechanism triggered by tumor growth. Altogether, our results identify a complex reciprocal relationship between adipocytes, immune cells, and the tumor, which may modulate the immune macroenvironment that promotes breast cancer development in obesity

Sepiolite and palygorskite-underpinned regulation of mRNA expression of pro-inflammatory cytokines as determined by a murine inflammation model

This paper shows that clay minerals, sepiolite and palygorskite collected from Torrejón El Rubio and Vallecas, Spain, respectively, altered the expression of four, namely, pro-inflammatory cytokines: interleukins IL-1 and IL-6, tumor necrosis factor (TNF-α), and interferon gamma (IFN-γ) as determined using a 12-O-tetradecanoylphorbol-13-acetate model for inflammation. Quantitative RT-PCR analyses after 4 and 24 h inflammatory stimuli showed that sepiolite or palygorskite brought about a reduction in mRNA expression. Sepiolite provoked the highest mRNA expression inhibition for all cytokines, except for TNF-α, and primarily after 4 h. Conversely, the anti-inflammatory effect for cytokine TNF-α was found to be true in the presence of palygorskite. Most notably, the significant reduction in mRNA expression of IL-1 registered just shortly after exposure pointed to that the anti-inflammatory effect may be important for modulation of the late inflammatory response. These clay minerals caused modifications in the mRNA expression of IL-1 and its receptor in endothelial cells and downstreaming inflammatory cascades resulting in the recruitment of neutrophils. In addition, polymorphonuclear peroxidase activity was severely reduced just after short exposure to either sepiolite or palygorskite. Evidence provided herein agree well with the idea that these clay minerals impaired neutrophils infiltration to inflamed skin, notwithstanding ear edema and deficient cell localization to skin coupled with such impairment may affect the later stages of inflammation.Peer reviewe

Sepiolite and palygorskite regulate gene expression of pro-inflammatory cytokines as determined by a murine inflammation model

Trabajo presentado a la XVI International Clay Conference (ICC), celebrada en Granada (España) del 17 al 21 de julio de 2017This presentation shows that sepiolite and palygorskite collected from Torrejón El Rubio and Vallecas, Spain modulate the inflammatory environment at its initial phase, consequently modify the initial and late phases of inflammation as well as edema in skin. At the same time, both clay minerals exert an effect on the recruitment of neutrophils to inflamed skin. These clay minerals, particularly sepiolite, cause a down-regulation of pro-inflammatory cytokine expressions, particularly those intrinsic of interleukin (IL-1) and tumor necrosis factor (TNF-α). The result is the modification of the local microenvironment at both stages of inflammation. We explain the effects of clay minerals on gene expression of pro-inflammatory cytokines because interactions between surface hydroxyl groups (≡Si-OH) and cytokines to form aggregates (e.g., ≡Si-O×××CO-NH-R), and subsequent stable organomineral derivatives (e.g., ≡Si-O-CO-NH-R); [1]]. Specifically, we propose binding of cytokine-gene promoter moieties at surface sites (type N or P) [2]. Taken together, results presented herein serve as the basis to support the potential use of these clay minerals as a therapeutic tool against inflammatory diseases. First, as determined by the 12-O-tetradecanoylphorbol-13-acetate model for inflammation [3], sepiolite and palygorskite altered the expression of four pro-inflammatory cytokines, namely, interleukins IL-1 andIL-6, TNF-α, and interferon gamma (IFN-γ). Second, Reverse Transcription Polymerase Chain Reaction (RT-PCR) analyses after 4 and 24 h inflammatory stimuli showed that either clay mineral brought about a reduction in mRNA expression. Sepiolite provoked the highest mRNA expression inhibition for all cytokines, except for TNF-α, and primarily after 4 h. Conversely, the anti-inflammatory effect for cytokine TNF-α was found to be true in the presence of palygorskite. Most notably was the significant reduction in mRNA expression of IL-1, registered just shortly after exposure, in keeping with the notion that the anti-inflammatory effect may be important for modulation of the late inflammatory response. Furthermore, sepiolite and palygorskite caused modifications in the mRNA expression of IL-1 and its receptor in endothelial cells and downstreaming inflammatory cascades resulting in the recruitment of neutrophils. Most remarkably, the activity of polymorphonuclear peroxidase was severely reduced just after short exposure to either clay mineral. In summary, sepiolite and palygorskite impaired neutrophils infiltration to inflamed skin [4], notwithstanding ear edema and deficient cell localization to skin coupled with such impairment may affect the later stages of inflammation [2,5].Peer reviewe

Analysis of RNA yield in extracellular vesicles isolated by membrane affinity column and differential ultracentrifugation.

Extracellular vesicles (EV) have attracted much attention as potential biomarkers due to their protein, RNA and other nucleic acid content. The most common method used for EV isolation is differential ultracentrifugation (DU), however given the DU technical difficulties, other more practical methods have surged, such as membrane-affinity column commercial kits. Here, we assessed one commercial kit in terms of EV recovery and EV-derived RNA yield and compared it with a DU protocol. Our data shows that the commercial kit preparation results in a lower count of EV-like structures and a reduced expression of EV markers when compared to DU samples. Thus, apparently suggesting that the commercial kit had a lower EV yield. However, these findings did not reflect on RNA yield, which was greater with the commercial kit, even after an enzymatic treatment with proteinase K and RNAse A. We conclude that the kit has a higher EV-derived RNA yield in comparison to our DU protocol, suggesting that it may be the method of choice for RNA sequencing purposes